The evidential value of dental calculus in the identification process.

DNA analysis-based identification is by far the gold standard in forensic genetics and it should be performed in every case involving human remains or unidentified bodies. Bones and teeth are the preferred source of human DNA for genetic analysis. However, there are cases where the nature of the proceedings and historical significance prevent the disruption of skeletal structure. The remains may also be heavily degraded. In such situations, forensic geneticists seek alternative sources of human DNA. Teeth calculus has proven to be a viable source of DNA for identification purposes. The aim of this study was to assess the concentration of human DNA in teeth calculus and evaluate the usefulness of teeth calculus as a DNA source in the identification process. Teeth calculus was collected from skeletons exhumed between 2021 and 2022 by the PBGOT (Polish Genetic Database of Victims of Totalitarianism) team from the former Stalag IID prisoner-of-war camp in Stargard. Genetic analyses included the determination of autosomal and Y-STR markers. The total concentration of human DNA was also evaluated in samples from teeth calculus and teeth taken from the same individuals. The pilot study included 22 skeletons with a sufficient amount of calculus for isolation (specified in the protocol). Samples were taken from the largest areas of calculus deposited on lingual surfaces of mandibular incisors. The prepared samples underwent DNA extraction. Our study demonstrated that teeth calculus is a source of human DNA for remains from the World War II period. The obtained DNA concentration allowed for the determination of STR markers. It was shown that teeth calculus contains human DNA in an amount suitable for preliminary identification analyses.

The last flight of F/O Tadeusz Stabrowski. Identification of the polish pilot.

The paper presents the process of identifying an unnamed soldier of the Polish armed forces in the west, whose remains were found in a nameless grave at the municipal cemetery in Le Crotoy in France. The Polish Genetic Database of Victims of Totalitarianism team carried out the research in cooperation with the Ministry of Culture and National Heritage. A comprehensive analysis of autosomal and Y-STR markers was performed. Historical, anthropological, and forensic examinations of the remains were also carried out. The items found with the remains were also examined. Identification based on DNA analysis made it possible to restore the identity of the Polish pilot who died on 11 March 1943 near the French coast, F/O Tadeusz Stabrowski. The airman regained his name in 2018, he was about 26 years old at the time of his death and left behind a grieving wife and son in the United Kingdom. The success of identifying the NN remains was guaranteed by the appointment of an interdisciplinary team consisting of specialists in archaeology, anthropology, history, forensic medicine and forensic genetics. The analysis of historical sources allowed to determine 4 missing airmen whose remains could have been buried in the cemetery in Le Crotoy. An interesting aspect of the research was the cooperation with history enthusiasts and fans of Polish aviation, thanks to which it was finally possible to narrow down the group of pilots sought and reach the family of Tadeusz Stabrowski, who submitted comparative material for research. This is the first case of establishing the identity of a Polish pilot killed in France. Many institutions have been involved in the project, including Polish Ministry of Culture and National Heritage (MDiKN), which partially funded the research.

Black Devils in Normandy-Identification of an Unknown Soldier Found in the Polish War Cemetery of Urville-Langannerie (France).

A paper dedicated to the identification of a Polish soldier from the 1st Armoured Division under the command of General Stanislaw Maczek, who fell in 1944 in Normandy, during World War II. The remains were found at the Urville-Langannerie Polish War Cemetery. A team from the Department of Forensic Genetics at the Pomeranian Medical University in Szczecin, commissioned by the Ministry of Culture Heritage and Sport, exhumed the remains in order to carry out genetic identification tests. A comprehensive anthropological analysis of the heavily degraded remains was carried out, and biological samples were secured for genetic testing. The identification of Jan Dusza is the first case of restoring the identity of an active combatant from the First Armoured Division. In the case analysis, the analysis of mitochondrial DNA in highly degraded biological material proved crucial. Genetic studies decided to reject the original historical hypothesis No. I at their preliminary stage. Regarding hypothesis No. II, a comprehensive genetic analysis of mitochondrial and autosomal DNA was carried out. Comparative material was obtained from the alleged victim's sister. Thanks to the analysis of kinship in the maternal line based on the mtDNA haplotype, it was possible to establish that the remains belong to Jan Dusza, who served in the Podhale Rifle Battalion, part of the Polish 1st Armoured Division. The research was co-financed by the Polish Ministry of Heritage and National Culture.

Is MPS always the answer? Use of two PCR-based methods for Y-chromosomal haplotyping in highly and moderately degraded bone material.

Forensic and population genetics often rely on Y-chromosomal studies. Whether it is a human identification case, trace evidence examination or phylogenetic analysis, a Y-STR haplotype is an important tool in the hands of law enforcement agencies. A common obstacle in achieving satisfactory results in all of the above mentioned circumstances, is low DNA quantity and quality within samples obtained. In this study we have examined Y-STR haplotypes in 75 bone material samples, coming from different time periods. For this purpose we have chosen YFiler Plus PCR Amplification Kit (ThermoFisher Scientific) and ForenSeq Signature DNA Prep Kit (Verogen Inc.), which use two different allele calling technologies - capillary electrophoresis and Massively Parallel Sequencing respectively. Full profiles were obtained from DNA extracts with as little as 0.1896 ng (Degradation Index 1.3) (ForenSeq) and 0.0591 ng (Degradation Index 26.8) (YFiler Plus) DNA input. The results that we present in this paper show differences in amplification rates between common markers in both kits. The differences strictly reflect mean amplicon length of markers. This, however, does not seem to influence Y-haplogroup estimation results noticeably. In one sample a discordance occurred between haplotypes obtained with both methods, where a 24 allele was called in DYS390 marker by capillary electrophoresis, while for the same sample in this locus a 23 allele was shown with MPS. A reason for this is yet to be investigated. The sequence analysis revealed a significant variation between isometric alleles, especially within repetitive regions of studied Y-STR markers.

Correlation Between Stem and Progenitor Cells Number and Immune Response in Patients After Allogeneic Kidney Transplant.

BACKGROUND Stem and progenitor cells are of great interest in all medical procedures involving tissue regeneration. There is a consensus that the use of stem cells after solid organ transplantation may play a role in tissue repair and in immunosuppression. The aim of this study was to determine possible relations between stem cell count and the immune response in a group of patients after kidney transplantation. MATERIAL AND METHODS The study was conducted on a group of 100 patients who underwent kidney transplantation. The following phenotypic markers of the studied cell subpopulations were adopted: Treg cells (CD3+CD4+CD25high), circulating hematopoietic cells (CD34+CD133+CD45+CD38-), and non-hematopoietic cells (Lin-CXCR4+CD133-CD45-). Cell subpopulations were assessed using LSRII flow cytometer (BD Biosciences, San Jose, CA, USA). RESULTS Positive correlation was observed between non-hematopoietic stem cells percentage and recipient's platelets count (P=0.04). Moreover, a higher percentage of non-hematopoietic cells was accompanied by lower numbers of B lymphocytes (P=0.03) and Treg cells (P=0.02). CONCLUSIONS Our study revealed significant associations between the intensity of ongoing immune response processes and tissue damage, and the release of stem and progenitor cells into circulation. These findings suggest their role in the stimulation of protective processes in terms of graft regeneration.

Genetic variation of 15 autosomal STRs in a population sample of Bedouins residing in the area of the Fourth Nile Cataract, Sudan.

ABSTRACT: The purpose of the paper was to report allelic frequencies of 15 autosomal STR markers (AmpFlSTR NGM PCR Amplification Kit) for Bedouin inhabitants in the area of the Fourth Nile Cataract in Sudan, and compute commonly used population and forensic biostatistical parameters. Buccal swabs were collected from 117 unrelated individuals. DNA was extracted using DNA QIAamp® DNA Mini Kit, and quantitated with Quantifiler Human Quantification Kit in a 7500 Real-Time PCR System. Amplification of 15 AmpFlSTR NGM PCR Kit loci was performed in PCR System 9700. Electrophoresis and typing were performed in 3130 Genetic Analyzer. Arlequin v3.5 software and PowerStats v1.2 spreadsheet were used for statistical calculations. The STR frequency distributions showed no deviations from HWE. The combined values of Matching Probability and Power of Exclusion are 1.77 × 10-18 and 0.9999996, respectively. The average observed heterozygosity over 15 loci is 0.8069. Five different allelic microvariants were found. A significant linkage disequilibrium was observed in five pairs of loci. A 15 STR population database has been established for Sudanese Bedouins. The systems studied have been shown to be useful tool for personal identification in this population.

Example of human individual identification from World War II gravesite.

This paper presents the procedure elaborated by our team which was applied to the mode of identification of Red Army soldiers who were taken as prisoners by the German Army during World War II and deceased in captivity. In the course of our search the unmarked burial of ten Soviet prisoners of war was found. Historical, anthropological and genetic research conducted by us led to the personal identification of nine of them, including two by means of DNA analysis.

Role of platelets in the modulation of kidney allograft recipients' immune systems.

BACKGROUND: Renal transplantation is the most effective method of treatment in end-stage renal disease. Chronic allograft rejection still remains a challenge for transplant physicians. Despite a growing amount of data regarding the role of platelets (PLT) in immunological processes, few reports have correlated number of platelets with transplanted kidney function. We aimed to evaluate the correlation between number of circulating platelets and number of immune system cells, including lymphocytes CD 4+, CD8+, lymphocytes B, monocytes, NK cells, and lymphocytes T reg in kidney transplant recipients with stable graft function. MATERIAL AND METHODS: We enrolled 100 kidney transplant recipients (ages 20-78 years) 10 month to 10 years after transplantation. The numbers of platelets (using standard procedure) and immune blood cells were evaluated using flow cytometry. Statistical analysis was performed with Spearman rank correlation. RESULTS: We found a negative correlation between number of platelets and number of lymphocytes T reg, and a positive correlation between platelet count and number of other examined immunocompetent cells. CONCLUSIONS: The number of PLT correlates with number of cells responsible for induction and effector mechanisms of acquired cellular response.

CAG repeat polymorphism in the androgen receptor gene in women with rheumatoid arthritis.

OBJECTIVE: Rheumatoid arthritis (RA) is the most common chronic, autoimmune, inflammatory disease, with a genetic and hormonal background. The prevalence of women among patients with RA suggests the important role of sex hormones in the pathogenesis of RA. We examined the association between CAG repeat polymorphism in the androgen receptor (AR) gene and susceptibility to RA and its clinical features in white women. METHODS: The study groups consisted of 325 female patients with RA and 238 female controls. CAG repeat polymorphism was determined using polymerase chain reaction and subsequent fragment analysis by capillary electrophoresis. RESULTS: The number of CAG repeats in patients did not differ from that of controls (22.1 ± 2.9 vs 21.9 ± 2.9, respectively; p = 0.26), but the presence of articular erosions was associated with a lower number of repeats in the shorter allele of patients with RA (20.4 ± 2.2 vs 21.2 ± 2.4; p = 0.031). When alleles with < 22 CAG were classified as short (S) and those with ≥ 22 CAG as long (L), the age at diagnosis of RA was lower in women with S-S genotype in comparison to combined S-L + L-L genotypes (43.0 ± 14.6 yrs vs 47.6 ± 12.5 yrs; p = 0.021). In patients with the L-L genotype, the frequency of erosive disease (OR 0.45, 95% CI 0.25-0.80, p = 0.0085) and extraarticular manifestations (OR 0.50, 95% CI 0.26-0.98, p = 0.047) was lower in comparison to carriers of the S allele. In multivariate analysis, the L-L genotype was an independent factor associated with a lower risk of erosions (OR 0.44, 95% CI 0.22-0.90, p = 0.024). CONCLUSION: The results suggest the association of short AR (CAG)(n) alleles with earlier onset and a more aggressive course of RA.

Lack of association between CAG repeat polymorphism in the androgen receptor gene and the outcome of rheumatoid arthritis treatment with leflunomide.

PURPOSE: Leflunomide (LEF) is a disease-modifying antirheumatic drug used for treating rheumatoid arthritis (RA) and the action of which may be modified by sex hormones. The aim of this study was to examine the association between CAG repeat polymorphism in the androgen receptor (AR) gene and the response to treatment with LEF in women with RA. METHODS: We studied 114 women diagnosed with RA and treated with LEF (20 mg daily). Follow-up was 12 months. CAG repeat polymorphism was determined using polymerase chain reaction (PCR) and subsequent fragment analysis by capillary electrophoresis. RESULTS: Analysis revealed no statistically significant associations between CAG repeat polymorphism in the AR gene and improvement of disease activity parameters: erythrocyte sedimentation rate, serum C-reactive protein, patient's global assessment of disease activity on a visual analog scale (VAS), disease activity score of 28 joints (DAS28), and swollen and tender joint count. CONCLUSION: Our results suggest no correlation between CAG repeat polymorphism in the AR gene and response to treatment with LEF in women with RA.

[Y-STR Poland--a database for evaluation of evidence value in forensic genetics]. / Y-STR Polska--baza danych do oceny wartosci dowodowej w genetyce sadowej.

The "Y-STR Poland" is a multicenter project, the aim of which is the construction of a widely available database of Y chromosome haplotypes determined in the Polish population in a range of sixteen loci in AmpFISTR Y-filer system. The database will be regularly updated and it will be used in assessment of evidence value in forensic genetics. The starting base "Y-STR Poland" contains 1600 Y-STR haplotypes and encompasses data collected in Lodz (two independent centers), Warsaw and Szczecin regions. The present report contains as an attachment the data in an Excel-type file, which serves as a tool in frequency determination of a given Y haplotype in the Polish population. The file will be updated on a regular basis along with updating the database, and will be freely available from www.genetyka-sadowa.pl.

Haptoglobin 2-1 phenotype predicts rapid growth of abdominal aortic aneurysms.

BACKGROUND: Haptoglobin (Hp) polymorphism is associated with the prevalence and clinical evolution of many inflammatory diseases and atherosclerosis. Circulating neutrophils and neutrophil-associated proteases are an important initial component of experimental abdominal aortic aneurysm (AAA) formation. Elastase and C-reactive protein (CRP) levels are elevated in patients with AAAs. This study assessed the relationship between AAA expansion and Hp phenotypes, neutrophil count, elastase, and CRP levels. METHODS: Eighty-three consecutive AAA patients underwent annual ultrasound scans. Three major Hp phenotypes (1-1, 2-1, and 2-2) were determined, and the neutrophil count, serum elastase, and high-sensitivity (hs) CRP levels were measured at the initial examination. After initial screening, patients were rescanned at 6- to 12-month intervals up to a period of 2 to 7 years. The mean yearly growth of the AAA largest transverse diameter was estimated for each group of Hp patients. The results are presented as median (interquartile range). RESULTS: Hp 2-1 patients had a significantly higher growth rate (3.69 [2.40] mm/y) of AAA compared with patients with Hp 2-2 (1.24 [0.79], P < .00001) and Hp 1-1 (1.45 [0.68], P = .00004). This association remained significant in the multivariate analysis. Elevated elastase serum activity was also evident in AAA patients with Hp 2-1 (0.119 [0.084] arbitrary units) in contrast to Hp 2-2 (0.064 [0.041], P < .00001) and Hp 1-1 (0.071 [0.040], P = .0006) patients. CRP serum levels (mg/L) were significantly higher in patients with Hp 2-1 (7.2 [7.1]) than in Hp 2-2 (3.4 [3.1], P = .0058) and Hp 1-1 (2.8 [4.1], P = .044). The neutrophil count was not significantly different among Hp groups. CONCLUSIONS: The Hp 2-1 phenotype showed a strong association with increased rates of the expansion of AAAs and may be a useful independent predictor of growth rate. Further large follow-up studies will be needed to investigate the pathomechanisms of association and the role of elastase and inflammation in the progression of AAA.

[Identification of the site of skeletal remains from World War II]. / Identyfikacja stanowiska szkieletowego z okresu II wojny swiatowej.

The objective of the present article is to show the model of processing in case of discovering a WW II grave with skeletal remains and to discuss the possibilities of personal identification of the victims. In many similar cases, it is impossible to employ genetic methods because of lack of samples for comparison. The shown procedure proved to be efficient in spite of the fact that more than 60 years passed since the date of death.

[The utility of the 16 Y-STRS markers in the central Poland population]. / Zastosowanie 16 markerów Y-STR w populacji Polski centralnej.

Haplotype and allele frequencies for the panel of 16 Y-chromosome STR loci, namely DYS19, DYS385, DYS389I, DYS389II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439, DYS448, DYS456, DYS458, DYS635 and Y GATA-H4 were determined in a population sample of 250 unrelated males from the central region of Poland. The 238 different haplotypes were identified, of which 227 haplotypes were unique, 10 were duplicated and one haplotype was shared between three unrelated men. The average gene diversity was 0.5995, combined gene diversity was 0.9998 and probability that two randomly chosen haplotypes are different in the population was 0.9996. Interpopulation comparisons revealed that Polish population is homogeneous within the three compared samples, while it differs statistically significantly from other population samples apart from Spanish population. This seems to be the first report showing the analysis of the molecular genetic distance between the population of Poland and other European and world populations for the panel of 16 Y-chromosome STR loci. The analyzed set of Y-STR markers is very useful in forensic casework to identify males and trace paternal lineages.

[Genetic identification of an osseous material dating from diverse historical periods]. / Identyfikacja genetyczna materialu kostnego z róznych okresów historycznych.

INTRODUCTION: Bad quality and slender amount of the material at our disposal during identity examination and ever growing number of expired and difficult identity cases compelled us to examine: 1) amplification grade and amount of human DNA in degraded biological material originating from diverse environments and historical periods; 20 operating mode in case of biological material from its preparation through isolation, amplification till interpretation of the achieved results; 3) usefulness of the accepted identification and amplification method while creating an identification procedure pattern in case of strongly decayed material. MATERIALS AND METHODS: The research material were samples isolated from: bone fragments collected from grave crypts, bone fragments collected during autopsies performed in the Forensic Medicine Institute PMA and during exhumations. DNA isolation was done by means of modified osseous DNA preparation method proposed by Zoledziewska and Dobosz. DNA amount was determined by means of ABI PRISM 7900HT Sequence Detection System apparatus with the use of Quantifiler Human DNA Qualification Set. The samples were amplified by means of AmpFlSTR SGM Plus set and were separated electrophoretically in the ABI PRISM 310 apparatus. CONCLUSIONS: 1. Environmental conditions like humidity, temperature, subsoil and microorganisms putrefying the body have the greatest influence on the preservation and DNA degradation grade. 2. Low amplification of long DNA fragments testifies that in the future the analysis of degraded DNA would be made with the use of sets based on the short fragments: ca. 100 pz. 3. Time that elapses since death is of less significance when we take into account tissues exposed to unfavourable, multifactoral environmental influence. 4. The method used and introduced modifications can be freely utilised in identification cases when the material is strongly decayed, coming from victims of mass disasters, terrorist attempts, genocide and natural cataclysms when the use of standard methods is impedimented or impossible.

[Examination over a foreign body left inside during an operation of a lumbar spine]. / Badanie nad cialem obcym pozostawionym w trakcie operacji kregoslupa ledzwiowego.

Procedure of an examination over a gauze swab removed during a repeated operation of a patient who had undergone earlier a neurosurgical operation of a lumbar spine has been described. The extracted gauze swab became an exhibit in a case at court for financial compensation. The court commissioned to explain "if it could be placed in another way than leaving it during the former surgical operation". The neurosurgeon who performed the former operation decidedly denied that the gauze swab in question could have been left by him. Hemogenetic and histologic examination gave basis to the statement that the exhibit was left inside during the former neurosurgical operation.

Genetic data on 10 STR loci a population of western Poland.

Allele frequencies for 10 STRs included in the AmpFlSTR SGM Plus kit were determined in a population sample of 668 unrelated individuals living in western Poland. All loci met Hardy-Weinberg expectations. Exact tests disequilibrium analysis revealed two departures from independence out of 45 pairwise comparisons. The combined matching probability (MP) and power of exclusion (PE) for all 15 loci are 2.56 x 10(-13) and 0.99996, respectively.